Expansion microscopy-based imaging of nuclear structures in cultured cells

Antoine Gaudreau-Lapierre; Kirk Mulatz; Jean-Claude Béïque; Laura Trinkle-Mulcahy

doi:10.1016/j.xpro.2021.100630

Expansion microscopy-based imaging of nuclear structures in cultured cells

STAR Protoc. 2021 Jun 26;2(3):100630. doi: 10.1016/j.xpro.2021.100630. eCollection 2021 Sep 17.

Authors

Antoine Gaudreau-Lapierre^{1

2}, Kirk Mulatz^{1

3}, Jean-Claude Béïque^{1

3}, Laura Trinkle-Mulcahy^{1

2}

Affiliations

¹ Department of Cellular and Molecular Medicine, University of Ottawa, Ottawa, ON K1H 8M5, Canada.
² Ottawa Institute of Systems Biology, University of Ottawa, Ottawa, ON K1H 8M5, Canada.
³ Brain and Mind Research Institute, University of Ottawa, Ottawa, ON K1H 8M5, Canada.

Abstract

Expansion microscopy is a sample preparation technique in which fixed and immunostained cells or tissues are embedded in a cross-linked network of swellable polyelectrolyte hydrogel that expands isotropically upon addition of deionized water. We utilize the X10 method for tenfold expansion of U2OS cells with concurrent DNA staining. A custom 3D-printed gel cutter and chambered slides minimize gel drift, facilitating analysis of the components of nuclear structures at nanoscale resolution by conventional microscopy or Airyscan confocal imaging. For complete information on the generation and use of this protocol, please refer to Do et al. (2020).

Keywords: Cell Biology; Microscopy.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Cell Line
Cell Nucleus / ultrastructure*
Microscopy / methods*
Reproducibility of Results
Software